Methods and compositions for modulating myofibroblast activities

ABSTRACT

The present invention discloses methods and compositions for treating or ameliorating a condition associated with increased or decreased myofibroblast activities and use thereof.

This application claims priority as a continuation to InternationalApplication No. PCT/US2012/042517, filed on Jun. 14, 2012, and entitled“METHODS AND COMPOSITIONS FOR MODULATING MYOFIBROBLAST ACTIVITIES,”which in turn claims priority to U.S. provisional application No.61/497,397 filed on Jun. 15, 2011, each of which is hereby incorporatedby reference herein in its entirety.

FIELD OF THE INVENTION

The present invention generally relates to methods and compositions fortreating or ameliorating a condition associated with increased ordecreased myofibroblast activities and use thereof.

BACKGROUND OF THE INVENTION

Excessive or deficient myofibroblast activity is associated with manydiseases and biological and medical processes. Such diseases includethose shown in Table 1*.

Tissue or Organ Activation/Proliferation Deletion or Damage SkinGranulation tissue Scleroderma; keloid; Dupuytren's contracture (72,213, 224); psoriasis (63) Pericyte Atherosclerosis and restenosis (149,159); Microancurysms, edema, hypertension (208) and hemorrhage (26, 239)Mouth Periodontal ligament Periodontal disease (136, 214) GingivalGingival hypertrophy secondary to drugs myofibroblasts (cyclosporine andDilantin)(135, 136, 212, 214, 216) Eye Orbital fibroblast Exophthalmos(proptosis) of Grave's disease (9, 221, 254) Retinal Proliferativevitreoretinopathy (253) myofibroblast Anterior capsule of Anteriorcapsular cataract (172, 217) Diabetic microaneurysm lens (26, 142, 239)Corneal Corneal scarring (184) myofibroblast Heart and pericardiumMyocardial fibrosis, atherosclerosis, and coronary artery restenosis(35, 149, 159. 258) Kidney Mesangial cell Proliferative and sclerosingglomerulonephritis Absence of glomerular (108, 184, 239) structure (141,234) Interstitial cell Renal tubulointerstitial fibrosis (171, 177, 198,239) Liver Perisinusoidal Fibrosis and cirrhosis (72, 88, 150) stellate(Ito cell) Ischemia reperfusion injury of hepatic transplantation (206)Necrotizing hepatitis (62) Pancreas Periacinal stellate Pancreaticfibrosis (4, 8) cell Lung Interstitial contractile Pulmonaryinterstitial fibrosis, idiopathic and Emphysema (25) cell drug-induced;sarcoidosis (105, 209, 214) Stomach and intestine Abnormal intestinalInterstitial cell of motility; hypertrophic Cajal pyloric stenosis;Hirschsprung's disease; Subepithelial Collagenous colitis; villousatrophy and crypt megacolon of piebaldism; myofibroblast hyperplasia;fibrosis of Crohn's disease (2, 86, idiopathic pseudo- 114, 131, 153)obstruction (33, 52, 115, Healing gastric ulcer 183, 212, 243, 248, 249)Brain Astrocyte Produce glial scar tissue (166) Human immunodeficiencyvirus-associated cognitive motor disease; spongiform encephalopathy(166) Breast Stromal Fibrocystic disease; desmoplastic reaction tomyofibroblast breast cancer (73, 214) Bone marrow Stromal Fibrosis inmyelodysplasia and neoplastic Aplastic anemia (182, 218) myofibroblastdiseases (182, 218) Joint Synoviocyte Rheumatoid pannus formation (11)*See Table 5 of Powell, et al., Myofibroblasts. I. Paracrine cellsimportant in health and disease. Am J Physiol Cell Physiol Jul. 1, 1999vol. 277 no 1 C1-C19, references shown in this table refers to thearticles cited in Powell, et al.

Thus far, there are no effective ways of treating or ameliorating manyof the conditions associated with excessive or deficient myofibroblastactivities.

Therefore, there is a need for methods and compositions for modulatingmyofibroblast activity.

The embodiments below address the above identified issues and needs.

SUMMARY OF THE INVENTION

In one aspect of the present invention, it is provided method fortreating or ameliorating a disorder associated with increased ordecreased myofibroblast activity, which method comprising modulating themyofibroblast level in a subject in need thereof by modulatingfibromodulin level in the subject. The subject can be a mammal, e.g., apatient or an animal.

In some embodiments of the method, modulating myofibroblast levelcomprises increasing fibromodulin level.

In some embodiments of the method, modulating myofibroblast levelcomprises decreasing fibromodulin level.

In some embodiments of the method, modulating fibromodulin level in thesubject comprises effectively modulating the level of fibromodulinactivity so as to modulate activities of a myofibroblast apoptosisinducing agent. Such myofibroblast apoptosis inducing agent can be,e.g., interleukin-1β (IL-1β).

In some embodiments of the method, modulating myofibroblast levelcomprises applying to the subject an effective amount of fibromodulinfor promoting myofibroblast differentiation and apoptosis therebyincreasing the myofibroblast level in the subject where early, but shortduration myofibroblast activity is effective for the disorder orinhibiting prolonged myofibroblast activity in the subject where thedisorder is associated with prolonged myofibroblast activity, whereinthe myofibroblast differentiation is induced by a myofibroblast inducingagent.

In some embodiments of the method, modulating myofibroblast levelcomprises blocking fibromodulin activity so as to prolong myofibroblastactivity.

In some embodiments of the method, blocking fibromodulin activitycomprises administering to the subject anti-sense oligonucleotides,antibodies, peptides that inhibit fibromodulin activities, andcombinations thereof.

In some embodiments of the method, the disorder is atherosclerosis,restenosis, cirrhosis, hemorrhage, microaneurysms, wounds with impairedcell motility, diabetic wounds, wounds with impaired tensile strength,wounds in patients on corticosteroids, wounds in olders, emphysema,interstitial lung diseases, asthma, Dupuytren's contracture, or anotherdisease in Table 1.

In some embodiments of the method, optionally in combination with any orall of the above embodiments, the myofibroblast differentiation inducingagent is TGF-β1.

In another aspect of the present invention, it is provided acomposition, which composition comprising an effective amount offibromodulin for promoting myofibroblast differentiation and apoptosisin a subject having a disorder associated with myofibroblast activity.

In some embodiments of the composition, the composition furthercomprises an excipient (e.g., a polymer or a formulation carrierdescribed below), wherein the formulation is a formulation for topical,transdermal, intradermal, or microneedle delivery.

In some embodiments of the composition, optionally in combination withany or all the above embodiments, the composition is included in amedical device comprising the composition.

In some embodiments, the medical device is a stent.

In a further aspect of the present invention, it is provided a method offorming a medical device, comprising applying a composition of thevarious above embodiments to the medical device. Such applying can be,e.g., forming a coating comprising the composition on the surface of themedical device. In some embodiments, the medical device is a stent.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 summarizes results of western blotting and quantitative RT-PCR(qRT-PCR) analyses which revealed that fibromodulin (FMOD) alone had nosignificant effect on rat dermal fibroblast (RDF) α-smooth muscle actin(α-SMA) expression. However, when in the presence of TGF-β1, FMODsignificantly enhances TGF-β1-induced α-SMA expression. Data arenormalized to untreated RDFs at time 0 (dash lines). N=3. *, P<0.05.

FIG. 2 shows results of an immunofluorescent (IF) staining study whichrevealed that FMOD, when combined with TGF-β1, significantly enhancedmyofibroblast differentiation and contractility in vitro. Formation ofstress fibers stained strongly for α-SMA are indicated by yellow arrows.Scale bar=100 μm.

FIG. 3 shows that FMOD+TGF-β1 significantly promotes TGF-β1-mediated RDFcontraction in collagen gel, while FMOD alone had minimal effects. N=6.

FIG. 4 shows the results of a PCR array assay which revealed thatexpression of α-SMA were significantly upregulated in FMOD-treated adultrat wounds at the early 3-day time-point, but reduced at the late 7- and14-day time-points. Gene expression is normalized to unwounded skin(dashed lines). N=9 wounds from 9 animals. **, P<0.01.

FIG. 5 shows that FMOD alone induces myofibroblast apoptosis aseffectively as IL-113, even in the presence of TGF-β1. N=6. *, P<0.05;**, P<0.01.

FIG. 6 shows results of PCR array assay which revealed that FMODsignificantly stimulated IL-1β expression in day 3 and 7 adult ratwounds. Gene expression is normalized to unwounded skin (dashed lines).N=9 wounds from 9 animals. **, P<0.01.

FIG. 7 shows that decreased α-SMA-positive myofibroblasts are found inFMOD-treated wounds at day 14 post injury. Scale bar=200 μm.

FIGS. 8A-8C show that FMOD reduces scar formation in adult rat woundsand increases tensile strength. At doses ≧0.4 mg/ml, histological (A)and quantitative analyses (B) demonstrate that FMOD significantlyreduces scar size of adult rat skin wounds compared to control at day 14post injury, while increasing tensile strength in a dose dependentmanner (C). Scale bar=25 μm.

DETAILED DESCRIPTION OF THE INVENTION

In one aspect of the present invention, it is provided method fortreating or ameliorating a disorder associated with increased ordecreased myofibroblast activity, which method comprising modulating themyofibroblast level in a subject in need thereof by modulatingfibromodulin level in the subject. The subject can be a mammal, e.g., apatient or an animal.

In some embodiments of the method, modulating myofibroblast levelcomprises increasing fibromodulin level. Increasing fibromodulin levelcan be achieved by endogenously increasing fibromodulin level via, e.g.,genetic engineering using a DNA or cDNA sequence expressing fibromodulinprotein or peptide. Genetic engineering technology is well establishedand can be readily achieved by a person of ordinary skill in the art.

In some embodiments, increasing fibromodulin level can be achieved byexogenously increasing fibromodulin level by, e.g., administering to asubject a fibromodulin protein or peptide. In some embodiments, suchfibromodulin protein or peptide can be included in a composition, whichcan be formulated into various formulations for different modes ofadministration. More detailed description of such formulations isprovided below.

In some embodiments of the method, modulating myofibroblast levelcomprises decreasing fibromodulin expression level. Decreasingfibromodulin expression level can be achieved by established method ofdecreasing expression level of a protein, e.g., gene knockout, genedeletion, etc. Such gene knockout or deletion methodologies are wellestablished and can be readily carried out by a person of ordinary skillin the art. Decreasing fibromodulin level can decrease myofibroblastdifferentiation induced by a myofibroblast inducing agent and apoptosisso as to prolong myofibroblast activity.

In some embodiments of the method, modulating fibromodulin level in thesubject comprises effectively modulating the level of fibromodulinactivity so as to modulate activities of a myofibroblast apoptosisinducing agent. Such myofibroblast apoptosis inducing agent can be,e.g., interleukin-1β (IL-1β).

In some embodiments of the method, modulating myofibroblast levelcomprises applying to the subject an effective amount of fibromodulinfor promoting myofibroblast differentiation and apoptosis therebyincreasing the myofibroblast level in the subject where early, but shortduration myofibroblast activity is effective for the disorder orinhibiting prolonged myofibroblast activity in the subject where thedisorder is associated with prolonged myofibroblast activity, whereinthe myofibroblast differentiation is induced by a myofibroblast inducingagent.

In some embodiments of the method, modulating myofibroblast levelcomprises blocking fibromodulin activity so as to prolong myofibroblastactivity. In some embodiments of the above methods, blockingfibromodulin activity comprises administering to the subject anti-senseoligonucleotides, antibodies, peptides that inhibit fibromodulinactivities, and combinations thereof. Medical conditions can benefitfrom (e.g., treated or ameliorated) prolonged myofibroblast activity. Anumber of examples of such medical conditions or disorders are shown inTable 1.

In some embodiments of the above methods, modulating myofibroblast levelcomprises applying to the subject an effective amount of fibromodulinfor promoting myofibroblast differentiation and apoptosis therebyincreasing the myofibroblast level in the subject where early, but shortduration myofibroblast activity is effective for the disorder orinhibiting prolonged myofibroblast activity in the subject where thedisorder is associated with prolonged myofibroblast activity. Suchdisorders include, e.g., wounds with impaired cell motility such asdiabetic wounds; wounds with impaired tensile strength such as wounds inpatients on corticosteroids; wounds in olders; emphysema; interstitiallung diseases; asthma; Dupuytren's contracture.

In some embodiments of the method, the disorder is atherosclerosis,restenosis, cirrhosis, hemorrhage, microaneurysms, wounds with impairedcell motility, diabetic wounds, wounds with impaired tensile strength,wounds in patients on corticosteroids, wounds in olders, emphysema,interstitial lung diseases, asthma, Dupuytren's contracture, or anotherdisease in Table 1.

In another aspect of the present invention, it is provided acomposition. The composition comprises an effective amount offibromodulin for promoting myofibroblast differentiation and apoptosisin a subject having a disorder associated with myofibroblast activity.

In some embodiments of the method, optionally in combination with any orall of the above embodiments, the myofibroblast differentiation inducingagent is TGF-β1.

In another aspect of the present invention, it is provided acomposition, which composition comprising an effective amount offibromodulin for promoting myofibroblast differentiation and apoptosisin a subject having a disorder associated with myofibroblast activity.

In some embodiments of the composition, the composition furthercomprises an excipient (e.g., a polymer or a formulation carrierdescribed below), wherein the formulation is a formulation for topical,transdermal, intradermal, or microneedle delivery.

In some embodiments of the composition, optionally in combination withany or all the above embodiments, the composition is included in amedical device comprising the composition.

In some embodiments, the medical device is a stent.

In a further aspect of the present invention, it is provided a method offorming a medical device, comprising applying a composition of thevarious above embodiments to the medical device. Such applying can be,e.g., forming a coating comprising the composition on the surface of themedical device. In some embodiments, the medical device is a stent.

As used herein, the term “myofibroblast differentiation inducing agent”shall mean any proteineous or small molecule agent capable of inducingmyofibroblast differentiation. An example of such myofibroblastdifferentiation inducing agents is transforming growth factor (TGF)-β1.

As used herein, the term “myofibroblast apoptosis inducing agent” shallmean any proteineous or small molecule agent capable of inducingmyofibroblast apoptosis. An example of such myofibroblast apoptosisinducing agents is interleukin-1β (IL-1β).

As used herein, the term “increased myofibroblast activity” shall meanthe level of myofibroblast activity exceeds the normal levelmyofibroblast activity in a subject (e.g., a human being), which iswithin the general knowledge of a medical practitioner. In some times,the term “increased myofibroblast activity” can be used interchangeablywith the term “excessive myofibroblast activity.”

As used herein, the term “decreased myofibroblast activity” shall meanthe level of myofibroblast activity falls below the normal levelmyofibroblast activity in a subject (e.g., a human being), which iswithin the general knowledge of a medical practitioner. In some times,the term “decreased myofibroblast activity” can be used interchangeablywith the term “insufficient myofibroblast activity.”

Myofibroblast Activity and Fibromodulin

Activity level of myofibroblast is related to the activity offibromodulin. Fibromodulin can promote myofibroblast differentiation andapoptosis. Fibromodulin addition in the presence of transforming growthfactor (TGF)-1β significantly promoted TGF-β1-mediated rat dermalfibroblasts (RDFs) α-smooth muscle actin (α-SMA, a wildly usedmyofibroblast marker) expression (FIG. 1). Remarkably, fibromodulin,when combined with TGF-β1, significantly enhanced myofibroblastdifferentiation and contractility in vitro (FIG. 2). Quantitative invitro collagen gel contraction studies confirmed increased contractilityof FMOD+TGF-β1 induced myofibroblasts relative to TGF-β1 alone (FIG. 3).In addition, expression of α-SMA were significantly upregulated inFMOD-treated adult rat wounds at the early 3-day time-point, but reducedat the late 7- and 14-day time-points (FIG. 4).

Notably, TGF-β1 completely blocked interleukin (IL)-β1-mediatedapoptosis in myofibroblasts (FIG. 5). Surprisingly, FMOD alone promotedmyofibroblast apoptosis as effectively as IL-1β, but more strikingly,FMOD administration promoted apoptosis even in the presence of TGF-1β(FIG. 5). Furthermore, FMOD significantly stimulated IL-1β expression inday 3 and 7 adult rat wounds (FIG. 6), which correlated functionallywith diminished α-SMA-positive myofibroblast numbers by day 14 postinjury (FIG. 7). Taken together, FMOD not only promotes TGF-β1-inducedmyofibroblast differentiation and contractility, but also acceleratesmyofibroblast clearance—resulting in lack of myofibroblast persistencein remodeling stage of FMOD-treated wounds.

As used herein, the term “fibromodulin” includes fibromodulin proteinand peptides. Examples of fibromodulin protein and peptides aredescribed in U.S. application Ser. No. 13/322,124, which is a nationalphase application of PCT/US2010/036262. Teachings in these applicationsare incorporated herein in their entirety by reference.

Formulation Carriers

The composition described herein may be administered to a subject inneed of treatment by a variety of routes of administration, includingorally and parenterally, (e.g., intravenously, subcutaneously orintramedullary), intranasally, as a suppository or using a “flash”formulation, i.e., allowing the medication to dissolve in the mouthwithout the need to use water, topically, intradermally, subcutaneouslyand/or administration via mucosal routes in liquid or solid form. Thepharmaceutical composition can be formulated into a variety of dosageforms, e.g., extract, pills, tablets, microparticles, capsules, oralliquid.

There may also be included as part of the composition pharmaceuticallycompatible binding agents, and/or adjuvant materials. The activematerials can also be mixed with other active materials includingantibiotics, antifungals, other virucidals and immunostimulants which donot impair the desired action and/or supplement the desired action.

In some embodiments, the composition can be formulated into aformulation for bone, which can include a carrier such as collagen,atelocollagen (collagen treated to remove the immunogenic ends),hydroxyapatite, and a polymer, which is further described below. Inthese embodiments, the formulation can comprise a porous matrix ormicrospheres made of a polymeric material, which is further describedbelow. In some embodiments, the polymer can be, e.g., polylactic acid orpolylactide (PLA), or poly(lactic acid-co-glycolic acid), or anotherbioabsorbable polymer.

In one embodiment, the mode of administration of the pharmaceuticalcomposition described herein is oral. Oral compositions generallyinclude an inert diluent or an edible carrier. They may be enclosed ingelatin capsules or compressed into tablets. For the purpose of oraltherapeutic administration, the aforesaid compounds may be incorporatedwith excipients and used in the form of tablets, troches, capsules,elixirs, suspensions, syrups, wafers, chewing gums and the like. Somevariation in dosage will necessarily occur, however, depending on thecondition of the subject being treated. These preparations shouldproduce a serum concentration of active ingredient of from about 0.01 nMto 1,000,000 nM, e.g., from about 0.2 to 40 μM. A preferredconcentration range is from 0.2 to 20 μM and most preferably about 1 to10 μM. However, the concentration of active ingredient in the drugcomposition itself depends on bioavailability of the drug and otherfactors known to those of skill in the art.

In another embodiment, the mode of administration of the pharmaceuticalcompositions described herein is topical or mucosal administration. Aspecifically preferred mode of mucosal administration is administrationvia female genital tract. Another preferred mode of mucosaladministration is rectal administration.

Various polymeric and/or non-polymeric materials can be used asadjuvants for enhancing mucoadhesiveness of the pharmaceuticalcomposition disclosed herein. The polymeric material suitable asadjuvants can be natural or synthetic polymers. Representative naturalpolymers include, for example, starch, chitosan, collagen, sugar,gelatin, pectin, alginate, karya gum, methylcellulose,carboxymethylcellulose, methylethylcellulose, andhydroxypropylcellulose. Representative synthetic polymers include, forexample, poly(acrylic acid), tragacanth, poly(methylvinylether-co-maleic anhydride), poly(ethylene oxide), carbopol,poly(vinyl pyrrolidine), poly(ethylene glycol), poly(vinyl alcohol),poly(hydroxyethylmethylacrylate), and polycarbophil. Other bioadhesivematerials available in the art of drug formulation can also be used(see, for example, Bioadhesion—Possibilities and Future Trends, Gurnyand Junginger, eds., 1990).

It is to be noted that dosage values also varies with the specificseverity of the disease condition to be alleviated. It is to be furtherunderstood that for any particular subject, specific dosage regimensshould be adjusted to the individual need and the professional judgmentof the person administering or supervising the administration of theaforesaid compositions. It is to be further understood that theconcentration ranges set forth herein are exemplary only and they do notlimit the scope or practice of the invention. The active ingredient maybe administered at once, or may be divided into a number of smallerdoses to be administered at varying intervals of time.

The composition may contain the following ingredients: a binder such asmicrocrystalline cellulose, gum tragacanth or gelatin; an excipient suchas starch or lactose, a disintegrating agent such as alginic acid,Primogel, corn starch and the like; a lubricant such as magnesiumstearate or Sterotes; a glidant such as colloidal silicon dioxide; and asweetening agent such as sucrose or saccharin or flavoring agent such aspeppermint, methyl salicylate, or orange flavoring may be added. Whenthe dosage unit form is a capsule, it may contain, in addition tomaterial of the above type, a liquid carrier such as fatty oil. Otherdosage unit forms may contain other various materials which modify thephysical form of the dosage unit, for example, as coatings. Thus tabletsor pills may be coated with sugar, shellac, or other enteric coatingagents. Materials used in preparing these various compositions should bepharmaceutically pure and non-toxic in the amounts used.

The solutions or suspensions may also include the following components:a sterile diluent such as water for injection, saline solution, fixedoils, polyethylene glycols, glycerine, propylene glycol or othersynthetic solvents; antibacterial agents such as benzyl alcohol ormethylparabens; antioxidants such as ascorbic acid or sodium bisulfite;chelating agents such as ethylenediaminetetraacetic acid; buffers suchas acetates, citrates or phosphates and agents for the adjustment oftonicity such as sodium chloride or dextrose. The parental preparationcan be enclosed in ampoules, disposable syringes or multiple dose vialsmade of glass or plastic.

The composition of the present invention can be prepared as formulationswith pharmaceutically acceptable carriers. Preferred are those carriersthat will protect the active compound against rapid elimination from thebody, such as a controlled release formulation, including implants andmicroencapsulated delivery systems. Biodegradable, biocompatablepolymers can be used, such as polyanhydrides, polyglycolic acid,collagen, and polylactic acid. Methods for preparation of suchformulations can be readily performed by one skilled in the art.

Liposomal suspensions (including liposomes targeted to infected cellswith monoclonal antibodies to viral antigens) are also preferred aspharmaceutically acceptable carriers. Methods for encapsulation orincorporation of compounds into liposomes are described by Cozzani, I.;Joni, G.; Bertoloni, G.; Milanesi, C.; Sicuro, T. Chem. Biol. Interact.53, 131-143 (1985) and by Jori, G.; Tomio, L.; Reddi, E.; Rossi, E. Br.J. Cancer 48, 307-309 (1983). These may also be prepared according tomethods known to those skilled in the art, for example, as described inU.S. Pat. No. 4,522,811 (which is incorporated herein by reference inits entirety). For example, liposome formulations may be prepared bydissolving appropriate lipid(s) (such as stearoyl phosphatidylethanolamine, stearoyl phosphatidyl choline, arachadoyl phosphatidylcholine, and cholesterol) in an inorganic solvent that is thenevaporated, leaving behind a thin film of dried lipid on the surface ofthe container. An aqueous solution of the active compound is thenintroduced into the container. The container is then swirled by hand tofree lipid material from the sides of the container and to disperselipid aggregates, thereby forming the liposomal suspension.

Other methods for encapsulating compounds within liposomes and targetingareas of the body are described by Sicuro, T.; Scarcelli, V.; Vigna, M.F.; Cozzani, I. Med. Biol. Environ. 15(1), 67-70 (1987) and Joni, G.;Reddi, E.; Cozzani, I.; Tomio, L. Br. J. Cancer, 53(5), 615-21 (1986).

The composition described herein may be administered in single (e.g.,once daily) or multiple doses or via constant infusion. The compounds ofthis invention may also be administered alone or in combination withpharmaceutically acceptable carriers, vehicles or diluents, in eithersingle or multiple doses. Suitable pharmaceutical carriers, vehicles anddiluents include inert solid diluents or fillers, sterile aqueoussolutions and various organic solvents. The pharmaceutical compositionsformed by combining the compounds of this invention and thepharmaceutically acceptable carriers, vehicles or diluents are thenreadily administered in a variety of dosage forms such as tablets,powders, lozenges, syrups, injectable solutions and the like. Thesepharmaceutical compositions can, if desired, contain additionalingredients such as flavorings, binders, excipients and the likeaccording to a specific dosage form.

Thus, for example, for purposes of oral administration, tabletscontaining various excipients such as sodium citrate, calcium carbonateand/or calcium phosphate may be employed along with variousdisintegrants such as starch, alginic acid and/or certain complexsilicates, together with binding agents such as polyvinylpyrrolidone,sucrose, gelatin and/or acacia. Additionally, lubricating agents such asmagnesium stearate, sodium lauryl sulfate and talc are often useful fortabletting purposes. Solid compositions of a similar type may also beemployed as fillers in soft and hard filled gelatin capsules. Preferredmaterials for this include lactose or milk sugar and high molecularweight polyethylene glycols. When aqueous suspensions or elixirs aredesired for oral administration, the active pharmaceutical agent thereinmay be combined with various sweetening or flavoring agents, coloringmatter or dyes and, if desired, emulsifying or suspending agents,together with diluents such as water, ethanol, propylene glycol,glycerin and/or combinations thereof.

For parenteral administration, solutions of the compounds of thisinvention in sesame or peanut oil, aqueous propylene glycol, or insterile aqueous solutions may be employed. Such aqueous solutions shouldbe suitably buffered if necessary and the liquid diluent first renderedisotonic with sufficient saline or glucose. These particular aqueoussolutions are especially suitable for intravenous, intramuscular,subcutaneous and intraperitoneal administration. In this connection, thesterile aqueous media employed are all readily available by standardtechniques known to those skilled in the art.

For intranasal administration or administration by inhalation, thecompounds of the invention are conveniently delivered in the form of asolution or suspension from a pump spray container that is squeezed orpumped by the patient or as an aerosol spray presentation from apressurized container or a nebulizer, with the use of a suitablepropellant, e.g., dichlorodifluoromethane, trichlorofluoromethane,dichlorotetrafluoroethane, carbon dioxide or other suitable gas. In thecase of a pressurized aerosol, the dosage unit may be determined byproviding a valve to deliver a metered amount. The pressurized containeror nebulizer may contain a solution or suspension of a compound of thisinvention. Capsules and cartridges (made, for example, from gelatin) foruse in an inhaler or insufflator may be formulated containing a powdermix of a compound or compounds of the invention and a suitable powderbase such as lactose or starch.

The composition described herein can be formulated alone or togetherwith the other agent in a single dosage form or in a separate dosageform. Methods of preparing various pharmaceutical formulations with acertain amount of active ingredient are known, or will be apparent inlight of this disclosure, to those skilled in this art. For examples ofmethods of preparing pharmaceutical formulations, see Remington'sPharmaceutical Sciences, Mack Publishing Company, Easton, Pa., 19thEdition (1995).

In some embodiments, the composition of the various embodimentsdisclosed above can be formulated into implants, scaffolds, patches,etc.

EXAMPLES

The embodiments of the present invention will be illustrated by thefollowing set forth examples. All parameters and data are not to beconstrued to unduly limit the scope of the embodiments of the invention.

Example 1

Using fibromodulin protein and peptide to accelerate myofibroblastdifferentiation and apoptosis or to inhibit prolonged myofibroblastactivity has been tested, and the results are positive (data not shown).

Expression of α-SMA were significantly upregulated in FMOD-treated adultrat wounds at the early 3-day time-point, but reduced at the late 7- and14-day time-point (FIG. 4). Notably, TGF-β1 completely blockedinterleukin (IL)-1β-mediated apoptosis in myofibroblasts (FIG. 5).Surprisingly, FMOD alone promoted myofibroblast apoptosis as effectivelyas IL-1β, but more strikingly, FMOD administration promoted apoptosiseven in the presence of TGF-1β (FIG. 5). Furthermore, FMOD significantlystimulated IL-1β expression in day 3 and 7 adult rat wounds (FIG. 6),which correlated functionally with diminished α-SMA-positivemyofibroblast numbers by day 14 post injury (FIG. 7). Taken together,FMOD not only promotes TGF-β1-induced myofibroblast differentiation andcontractility, but also accelerates myofibroblast clearance—resulting inlack of myofibroblast persistence in remodeling stage of FMOD-treatedwounds. As the result of this, FMOD treated adult rat primary closurewounds healed with significant smaller scar accompanied with moreorganized collagen architecture and stronger tensile strength (FIG. 8).

While particular embodiments of the present invention have been shownand described, it will be obvious to those skilled in the art thatchanges and modifications can be made without departing from thisinvention in its broader aspects. Therefore, the appended claims are toencompass within their scope all such changes and modifications as fallwithin the true spirit and scope of this invention.

We claim:
 1. A method for treating or ameliorating a disorder associatedwith increased or decreased myofibroblast activity, comprising:modulating the myofibroblast level in a subject in need thereof bymodulating fibromodulin level in the subject by increasing fibromodulinlevel or by decreasing fibromodulin level, or modulating myofibroblastdifferentiation and myofibroblast apoptosis, wherein increasingfibromodulin level comprises applying to the subject an effective amountof fibromodulin for promoting the myofibroblast differentiation andmyofibroblast apoptosis thereby increasing the myofibroblast level inthe subject where early, but short duration myofibroblast activity iseffective for the disorder or inhibiting prolonged myofibroblastactivity in the subject where the disorder is associated with prolongedmyofibroblast activity, wherein the myofibroblast differentiation isincreased by increasing the level of a myofibroblast inducing agent andthe myofibroblast apoptosis is induced by a myofibroblast apoptosisinducing agent, and wherein modulating fibromodulin level in the subjectcomprises effectively modulating the level of fibromodulin activities soas to modulate the myofibroblast apoptosis inducing agent.
 2. The methodof claim 1, wherein modulating myofibroblast level comprises blockingfibromodulin activity so as to prolong myofibroblast activity.
 3. Themethod of claim 2, wherein blocking fibromodulin activity comprisesadministering to the subject anti-sense oligonucleotides, antibodies,peptides that inhibit fibromodulin activities, and combinations thereof.4. The method of claim 1, wherein the disorder is atherosclerosis,restenosis, cirrhosis, hemorrhage, microaneurysms, wounds with impairedcell motility, diabetic wounds, wounds with impaired tensile strength,wounds in patients on corticosteroids, wounds in elders, emphysema,interstitial lung diseases, asthma, Dupuytren's contracture, or anotherdisease selected from the group consisting of scleroderma, keloid,Dupuytren's contracture, psoriasis, atherosclerosis and restenosis,hypertension, periodontal disease, gingival hypertrophy secondary todrugs (cyclosporine and Dilantin), exophthalmos (proptosis) of Grave'sdisease, Proliferative vitreoretinopathy, anterior capsular cataract,corneal scarring, myocardial fibrosis, atherosclerosis, coronary arteryrestenosis, proliferative and sclerosing glomerulonephritis, renaltubulointerstitial fibrosis, fibrosis, cirrhosis, ischemia reperfusioninjury of hepatic transplantation, necrotizing hepatitis, pancreaticfibrosis, pulmonary interstitial fibrosis, idiopathic and drug-induced,sarcoidosis, collagenous colitis; villous atrophy and crypt hyperplasia;fibrosis of Crohn's disease, healing gastric ulcer, produce glial scartissue, fibrocystic disease, desmoplastic reaction to breast cancer,fibrosis in myelodysplasia and neoplastic diseases, rheumatoid pannusformation, Microancurysms, edema, hemorrhage, diabetic microaneurysm,absence of glomerular structure, emphysema, abnormal intestinalmotility, hypertrophic pyloric stenosis; Hirschsprung's disease;megacolon of piebaldism; idiopathic pseudo-obstruction, humanimmunodeficiency virus-associated cognitive motor disease; spongiformencephalopathy, and aplastic anemia.
 5. The method of claim 1, whereinthe myofibroblast differentiation inducing agent is TGF-β1.
 6. Themethod of claim 1, wherein the myofibroblast apoptosis inducing agent isinterleukin-1β (IL-1β).
 7. The method of claim 1, wherein the subject isa patient.